Ceftolozane pharmaceutical compositions

ABSTRACT

Pharmaceutical compositions can include an amount of sodium chloride effective to stabilize ceftolozane in an antibiotic formulation.

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 61/792,092, filed Mar. 15, 2013, and U.S. Provisional PatentApplication No. 61/793,007, filed Mar. 15, 2013, both of which areincorporated herein in their entirety.

TECHNICAL FIELD

This disclosure relates to pharmaceutical compositions comprisingceftolozane.

BACKGROUND

Ceftolozane is a cephalosporin antibacterial agent, also referred to asCXA-101, FR264205, or by chemical names such as(6R,7R)-3-[(5-amino-4-{[(2-aminoethyl)carbamoyl]amino}-1-methyl-1H-pyrazol-2-ium-2-yl)methyl]-7-({(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-[(1-carboxy-1-methylethoxy)imino]acetyl}amino)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate,and7β-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-{3-amino-4-[3-(2-aminoethyl)ureido]-2-methyl-1-pyrazolio}methyl-3-cephem-4-carboxylate.

The prior art describes a variety of ceftolozane salts. For example,U.S. Pat. No. 7,129,232 discloses ceftolozane hydrogen sulfate saltamong other salts “with a base or an acid addition salt such as a saltwith an inorganic base, for example, an alkali metal salt [e.g., sodiumsalt, potassium salt, etc.], an alkaline earth metal salt [e.g., calciumsalt, magnesium salt, etc.], an ammonium salt; a salt with an organicbase, for example, an organic amine salt [e.g., trimethylamine salt,triethylamine salt, pyridine salt, picoline salt, ethanolamine salt,triethanolamine salt, dicyclohexylamine salt,N,N′-dibenzylethylenediamine salt, etc.]; an inorganic acid additionsalt [e.g., hydrochloride, hydrobromide, sulfate, hydrogen sulfate,phosphate, etc.]; an organic carboxylic or sulfonic acid addition salt[e.g., formate, acetate, trifluoroacetate, maleate, tartrate, citrate,fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.];and a salt with a basic or acidic amino acid [e.g., arginine, asparticacid, glutamic acid, etc.].” Ceftolozane sulfate is a pharmaceuticallyacceptable ceftolozane salt of formula (I) that can be formulated forintravenous administration or infusion.

Certain pharmaceutical compositions containing ceftolozane are useful asantibiotics for the treatment of certain serious infections, includingserious complicated intra-abdominal infections and complicated urinarytract infections. The ceftolozane pharmaceutical compositions can beadministered as intravenous antibacterial agents to treat theseinfections. The antibacterial activity of ceftolozane is believed toresult from its interaction with penicillin binding proteins (PBPs) toinhibit the biosynthesis of the bacterial cell wall which acts to stopbacterial replication. Antibacterial pharmaceutical compositions caninclude a therapeutically effective unit dose of a pharmaceuticallyacceptable salt of ceftolozane formulated for intravenousadministration.

As disclosed herein, ceftolozane is unstable in certain lyophilizedpharmaceutical compositions initially evaluated for intravenousadministration. In particular, a decrease in ceftolozane purity and theformation of multiple additional related substances were detected incertain initial ceftolozane pharmaceutical compositions by peaks in highpurity liquid chromatography (HPLC) after stability testing. Thistesting pointed to the need to develop novel ceftolozane formulationsproviding increased ceftolozane stability. U.S. Pat. No. 7,129,232discloses that “auxiliary substances” such as “stabilizing agents . . .and other commonly used additives” may be included in pharmaceuticalcompositions comprising ceftolozane or many other cephalosporincompounds “if needed.” However, the disclosure does not disclose areduction in ceftolozane purity in pharmaceutical compositionscontaining ceftolozane sulfate, or the formation of additional relatedsubstances observed during stability testing. Nor does the disclosureprovide guidance on the formation of ceftolozane pharmaceuticalcompositions to increase ceftolozane purity during stability testing orcontrol the relative amounts of ceftolozane related substances detectedby HPLC peak formation during stability testing.

In view of the above, there is a need for pharmaceutical preparationscontaining ceftolozane compounds having improved ceftolozane stability.

SUMMARY

Ceftolozane sulfate is stabilized in solid phase pharmaceuticalcompositions by incorporation of an effective amount of an inorganicsalt stabilizing agent, in particular 125 to 500 mg (more specifically480 to 500 mg) of sodium chloride per gram of ceftolozane active. Theinvention is based in part on the surprising discovery that ceftolozanepharmaceutical compositions comprising 125 to 500 mg (more specifically480 to 500 mg) of sodium chloride per 1000 mg of ceftolozane activedemonstrate improved ceftolozane purity and chemical stability comparedpharmaceutical compositions comprising ceftolozane with comparativelyless sodium chloride. For example, the disclosed pharmaceuticalcompositions have an improved stability as a decrease in the rate ofceftolozane purity and/or a decrease in the rate of formation ofsubstances characterized by HPLC peaks 1 and 7 identified during a 7-daystability study in Example 3. The disclosed ceftolozane pharmaceuticalcompositions comprise a stabilizing amount of sodium chloride (e.g., 125to 500 mg of sodium chloride [more specifically, 480 to 500 mg) per 1000mg of ceftolozane active). Certain preferred compositions demonstrateimproved ceftolozane purity (e.g., Table 3 in FIG. 5A) and chemicalstability (e.g., with respect to the composition of HPLC peak 1 in Table4, FIG. 6A) compared with pharmaceutical compositions comprisingceftolozane with comparatively less sodium chloride. For example, thedisclosed pharmaceutical compositions typically comprise less than about4% total impurity after being stored (e.g., in a sealed container) forseven days at 60° C., as determined by HPLC using a Develosil columnODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodiumperchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/minflow rate and oven temperature of 45° C. Alternatively, the disclosedpharmaceutical compositions comprise less than about 2% of the impurityrepresented by Peak 1 after being stored (e.g., in a sealed container)for seven days at 60° C., as determined by HPLC using a Develosil columnODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodiumperchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/minflow rate and oven temperature of 45° C., where Peak 1 has a retentiontime relative to ceftolozane of about 0.1.

Accordingly, preferred pharmaceutical antibiotic compositions caninclude ceftolozane sulfate and stabilizing amount of sodium chloride(e.g., 125 to 500 mg more specifically 480 to 500 mg of sodium chlorideand 1,000 mg ceftolozane active) in a unit dosage form (e.g., powder ina vial). The unit dosage form can be dissolved with a pharmaceuticallyacceptable carrier, and then intravenously administered.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B are chromatograms of CXA-101 ceftolozane drug substanceobtained from the lyophilization process of Example 1. The chromatogramswere obtained according to the analytical method described in Example 2.

FIG. 2 is a diagram of a lyophilization process for the ceftolozaneobtained according to the process described in Example 1.

FIG. 3 is a table (Table 1) of peaks for the ceftolozane prepared by thelyophilization process in Example 1 obtained by HPLC according to theanalytical method of Example 2.

FIG. 4 is a table (Table 2) showing the composition of variousadditional ceftolozane pharmaceutical compositions in which the sodiumchloride content is varied.

FIG. 5A is a table (Table 3) showing the total purity of ceftolozane inthe pharmaceutical compositions of FIG. 4, as measured by HPLC peak areaaccording to the analytical method of described in Example 2.

FIG. 5B is a graph showing the total purity of certain pharmaceuticalcompositions disclosed in FIG. 4, as measured by HPLC peak area.

FIG. 6A is a table (Table 4) showing the amount of material fromcharacteristic peak 1 in the pharmaceutical compositions of FIG. 4, asmeasured by HPLC peak area according to the analytical method ofdescribed in Example 2.

FIG. 6B is a graph showing the amount of material from characteristicpeak 1 in the pharmaceutical compositions of FIG. 4, as measured by HPLCpeak area according to the analytical method of described in Example 2.

FIG. 7A is a table (Table 5) showing the amount of material fromcharacteristic peak 3 in the pharmaceutical compositions of FIG. 4, asmeasured by HPLC peak area according to the analytical method ofdescribed in Example 2.

FIG. 7B is a graph showing the amount of material from characteristicpeak 3 in the pharmaceutical compositions of FIG. 4, as measured by HPLCpeak area according to the analytical method of described in Example 2.

FIG. 8A is a table (Table 6) showing the amount of material fromcharacteristic peak 7 in the pharmaceutical compositions of FIG. 4, asmeasured by HPLC peak area according to the analytical method ofdescribed in Example 2.

FIG. 8B is a graph showing the amount of material from characteristicpeak 7 in the pharmaceutical compositions of FIG. 4, as measured by HPLCpeak area according to the analytical method of described in Example 2.

FIG. 9A is a table (Table 7) showing the formulation composition of theCo-Lyo-Combo Drug Product used in Example 4A.

FIG. 9B is a table (Table 8) showing impurity concentrations at timezero, one month and three months at 25° C./60% relative humidity of theCo-Lyo-Combo Drug Product.

FIG. 9C is a table (Table 9) showing impurity concentrations at timezero, one month and three months at 40° C./75% relative humidity of theCo-Lyo-Combo Drug Product.

FIG. 10A is a table (Table 10) showing the formulation composition ofthe Blend Drug Product used in Example 4B.

FIG. 10B is a table (Table 11) showing impurity concentrations at timezero, one month and three months at 25° C./60% relative humidity of theBlend Drug Product used in Example 4B.

FIG. 10C is a table (Table 12) showing impurity concentrations at timezero, one month and three months at 40° C./75% relative humidity of theBlend Drug Product used in Example 4B.

FIG. 11 is a table (Table 13) showing the composition of variousceftolozane pharmaceutical compositions in which the sodium chlorideconcentration is varied.

FIG. 12 is a table (Table 14) showing the purity of Ceftolozane inCXA-201 Compositions with varying amounts of sodium from sodium chlorideat time zero, 1 day, 3 days and 7 days at 30° C. and 60° C.

FIG. 13 is a table (Table 15) showing the HPLC area of Impurity of Peak1 in CXA-201 Compositions with varying amounts of sodium from sodiumchloride at time zero, 1 day, 3 days and 7 days at 30° C. and at 60° C.

FIG. 14 is a table (Table 16) showing the HPLC area of the Impurity atRRT 0.43 and Impurity Peak 3 in CXA-201 Compositions with varyingamounts of sodium from sodium chloride at time zero, 1 day, 3 days and 7days at 30° C. and at 60° C.

FIG. 15 is a table (Table 17) showing the HPLC area of Impurity of Peak7 in CXA-201 Compositions with varying amounts of sodium from sodiumchloride at time zero, 1 day, 3 days and 7 days at 30° C. and 60° C.

FIG. 16 is a table (Table 18) showing the finished drug product unitcomposition of ceftolozane/tazolactam.

FIG. 17 is a table (Table 19) showing the primary container closuresystem for the ceftolozane/tazobactam unit product.

DETAILED DESCRIPTION

Ceftolozane sulfate can be stabilized in solid phase pharmaceuticalcompositions by incorporation of an effective amount of an inorganicsalt stabilizing agent, in particular 125 to 500 mg (more specifically480 to 500 mg) of sodium chloride per 1,000 mg of ceftolozane active ina liquid state, followed by conversion of the resulting solution to asolid.

In particular, pharmaceutical compositions comprising ceftolozane andstabilizing amount of sodium chloride can be obtained by lyophilizationof a solution comprising a stabilizing-effective amount of sodiumchloride and ceftolozane sulfate. Alternatively, they can be obtained byother methods. As is known to those skilled in the art, lyophilizationis a process of freeze-drying in which water is sublimed from a frozensolution of one or more solutes. Specific methods of lyophilization aredescribed in Remington's Pharmaceutical Sciences, Chapter 84, page 1565,Eighteenth Edition, A. R. Gennaro, (Mack Publishing Co., Easton, Pa.,1990). A pharmaceutical composition comprising ceftolozane can beprepared by adding a stabilizing amount of sodium chloride in a fixedratio to ceftolozane in an aqueous solution prior to lyophilization orspray drying, then lyophilizing or spray drying the solution to obtain alyophilized or spray dried composition comprising sodium chloride andceftolozane.

When obtained by lyophilization, the pharmaceutical antibioticcompositions can include ceftolozane sulfate obtained by a processcomprising the steps of lyophilizing an aqueous solution containingceftolozane and a stabilizing amount of sodium chloride, where thestabilizing amount of sodium chloride is about 125 to 500 mg (morespecifically 480 to 500 mg) of sodium chloride per 1000 mg ceftolozaneactive in the aqueous solution prior to lyophilization or spray drying.The pharmaceutical compositions comprising ceftolozane disclosed hereincan be prepared by adding 125 to 500 mg sodium chloride (morespecifically 480 to 500 mg) per 1000 mg of ceftolozane prior tolyophilization or spray drying. For example, the pharmaceuticalcompositions can be obtained by a method comprising the steps of addinga stabilizing amount (e.g., 125 to 1000 mg sodium chloride [morespecifically 480 to 500 mg ] per 1000 mg of ceftolozane active) ofsodium chloride to ceftolozane followed by lyophilizing or spray dryingthe composition comprising the sodium chloride and ceftolozane. In oneaspect (e.g., Example 1), ceftolozane and a stabilizing amount of sodiumchloride can be dissolved in an aqueous solution that can be lyophilizedto obtain a ceftolozane pharmaceutical composition.

The pharmaceutical compositions may comprise other additional componentsincluding stabilizers, pH adjusting additives (e.g., buffers) and thelike. Non-limiting examples of these additives include sodium chloride,citric acid and L-arginine. For example, the pharmaceutical antibioticcompositions can include ceftolozane sulfate obtained by a processcomprising the steps of lyophilizing or spray drying an aqueous solutioncontaining ceftolozane sulfate with a stabilizing amount of sodiumchloride (e.g., 125 to 500 mg of sodium chloride [more specifically 480to 500 mg] per 1,000 mg ceftolozane active), with L-arginine and/orcitric acid in the aqueous solution prior to lyophilization. The use ofsodium chloride results in greater ceftolozane stability, while anamount of L-arginine can be used that is effective to adjust pH and toincrease the solubility of ceftolozane, and citric acid can be includedin an amount effective to reduce or prevent discoloration of theproduct, due to its ability to chelate metal ions. The aqueous solutioncan optionally be subsequently lyophilized or spray dried to obtain astabilized lyophilized ceftolozane sulfate composition comprisingceftolozane sulfate, sodium chloride, L-arginine and citric acid.

The pharmaceutical composition can also be a Ceftolozane/Tazobactam forInjection Drug Product, 1000 mg/500 mg. It is presented as a combinationof two sterile active powders in a single vial intended forreconstitution and intravenous infusion.

The drug product is first prepared by converting ceftolozane sulfatedrug substance to a sterile drug product intermediate (DPI) powder withexcipients citric acid, sodium chloride and L-arginine. This is commonlydone by lyophilization, as described above. Tazobactam sodium drugsubstance is presented as a sterile powder without any excipients. Thetazobactam sodium drug substance is typically lyophilized, spray driedor provided as a crystalline material. The drug product is then preparedby aseptically filling the two powders (e.g., the two separatelylyophilized drug powders) sequentially into a single vial.

Each vial of ceftolozane/tazobactam for injection contains approximately2255 mg ceftolozane sterile DPI powder that contains 1147 mg ceftolozanesulfate, which is equivalent to 1000 mg ceftolozane free base, as wellas approximately 537 mg tazobactam sodium sterile drug substance,equivalent to 500 mg tazobactam free acid. At the time ofadministration, the vial is reconstituted with 10 mL vehicle, sterile 5%Dextrose Injection USP, Water for Injection or 0.9% Sodium ChlorideInjection USP, then the vial contents further diluted in an infusion bagof 0.9% Sodium Chloride Injection USP or 5% Dextrose Injection USP, foradministration. The constituents are shown in Table 18, FIG. 16.

The primary container-closure system is a Type 1 20 mL molded glass vialwith 20 mm neck finish. The vial is sealed by a 20 mm rubber stopper and20 mm plastic flip-cap seal with aluminum ferrule. The primarycontain-closure system used for the ceftolozane/tazobactam unit productis summarized in Table 19, FIG. 17.

In other embodiments, pharmaceutical compositions comprising ceftolozanecan be obtained by methods that include the steps of: (1) adding astabilizing amount of sodium chloride to ceftolozane optionally followedby co-lyophilizing or spray drying the ceftolozane and sodium chloride;and (2) combining the product of step (1) with other components. Forexample, the product of step (1) can be combined with a β-lactamaseinhibitor, such as tazobactam (CAS #: 89786-04-9), avibactam (CAS#1192500-31-4), Sulbactam (CAS #68373-14-8) and/or clavulanate (CAS#58001-44-8). The beta lactamase inhibitor can be included in acrystalline or amorpous form, such as a lyophilized tazobactam orcrystalline tazobactam (e.g., U.S. Pat. Nos. 8,476,425 and 5,763,603) toobtain the pharmaceutical composition.

Ceftolozane compositions having 50-481 mg of sodium chloride per 1,000mg ceftolozane active were prepared as described in Table 2 (FIG. 4) andtested for stability as described in Example 3. Ceftolozane was morestable in compositions containing at least 125 mg of sodium chloride per1,000 mg of ceftolozane active, as measured by high performance liquidchromatography (HPLC) analysis by detecting the ratio of peak areasobtained for ceftolozane compared to peaks for other substances. Unlessotherwise indicated, HPLC measurements reported herein are obtainedusing a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobilephase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v)at a 1.0 mL/min flow rate and oven temperature of 45° C.

During the stability test of Example 3, ceftolozane samples containing125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozaneactive showed a decrease in ceftolozane total purity measured by HPLCthat was at least about 35% less than reductions in ceftolozane totalpurity observed for formulations containing 50 mg or 75 mg sodiumchloride per 1,000 mg ceftolozane active. Thus, ceftolozane compositionshaving at least 125 mg or more sodium chloride relative to the fixedamount of ceftolozane were about 35-90% more stable than comparableceftolozane compositions having less than 125 mg sodium chloride (e.g.,the % decrease in ceftolozane for the sample containing 75 mg sodiumchloride was about 35% greater than the comparable % decrease inceftolozane for the sample containing 190 mg sodium chloride). Inaddition, samples obtained from ceftolozane compositions containing 125mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozaneactive showed a decrease in ceftolozane that was up to about 90% lessthan reductions in ceftolozane observed for formulations containing 50mg or 75 mg sodium chloride per (e.g., the % decrease in ceftolozane forthe sample containing 50 mg sodium chloride was about 90% greater thanthe comparable % decrease in ceftolozane for the sample containing 481mg sodium chloride).

The ceftolozane sodium-stabilized compositions having 125 mg or moresodium chloride relative to the fixed amount of 1,000 mg ceftolozaneactive also had lower quantities of additional substances identified bypeaks 1 and 7 having characteristic retention times measured by HPLC(see Table 1, FIG. 3, indicating retention times of about 0.1 for peak 1and about 1.3 for peak 7 relative to ceftolozane measured according tothe HPLC method of Example 2). In particular, these sodium chloridestabilized ceftolozane compositions were characterized by about 37-94%less of the material of peak 1 and about 38-306% less of the material ofpeak 7 (measured by corresponding HPLC peak areas) than comparableceftolozane compositions having less than 125 mg sodium chloride (e.g.,see 7-day stability study in Example 3). Referring to the data in Table4 (FIGS. 6A and 6B), the amount of the composition of peak 1 (measuredby HPLC according to Example 2) was measured by the % increase in thepeak 1 HPLC peak during the 7-day stability test of Example 3.

In particular, samples containing 125 mg, 190 mg and 481 mg of sodiumchloride per 1,000 mg of ceftolozane active showed at least a 37%reduction in the amount of the peak 1 composition observed for theseformulations containing at least 125 mg sodium chloride per 1,000 mgceftolozane active, compared to the compositions with 50 mg or 75 mgsodium chloride per 1,000 mg of ceftolozane active (e.g., the % increasein peak 1 for the sample containing 75 mg sodium chloride was about 37%greater than the comparable % decrease in ceftolozane for the samplecontaining 190 mg sodium chloride). In addition, compositions containing125 mg, 190 mg and 481 mg of sodium chloride per 1,000 mg of ceftolozaneactive showed up to a 94% reduction in the amount of the peak 1composition observed for these formulations containing at least 125 mgsodium chloride per 1,000 mg ceftolozane active, compared to thecompositions with 50 mg or 75 mg sodium chloride per 1,000 mg ofceftolozane active (e.g., the % increase in peak 1 for the samplecontaining 50 mg sodium chloride was about 94% greater than thecomparable % decrease in ceftolozane for the sample containing 481 mgsodium chloride).

Referring to the data in Table 5 (FIGS. 7A and 7B), the amount of thecomposition of peak 3 (measured by HPLC according to Example 2) wasmeasured by the % increase in the peak 3 HPLC peak during the 7-daystability test of Example 3.

Referring to the data in Table 6 (FIGS. 8A and 8B), the amount of thecomposition of peak 7 (measured by HPLC according to Example 2) wasmeasured by the % increase in the peak 7 HPLC peak during the 7-daystability test of Example 3.

Pharmaceutical compositions comprising ceftolozane can be formulated totreat infections by parenteral administration (including subcutaneous,intramuscular, and intravenous) administration. Pharmaceuticalcompositions may additionally comprise excipients, stabilizers, pHadjusting additives (e.g., buffers) and the like. Non-limiting examplesof these additives include sodium chloride, citric acid and L-arginine.For example, the use of sodium chloride results in greater stability;L-arginine is used to adjust pH and to increase the solubility ofceftolozane; and citric acid is used to prevent discoloration of theproduct, due to its ability to chelate metal ions. In one particularembodiment, the pharmaceutical compositions described herein areformulated for administration by intravenous injection or infusion.Pharmaceutical antibiotic compositions can include ceftolozane sulfateand stabilizing amount of sodium chloride (e.g., 125 to 500 mg (morespecifically 480-500 mg) of sodium chloride per 1,000 mg ceftolozaneactive), optionally in a lyophilized or spray dried unit dosage form(e.g., powder in a vial). The unit dosage form can be dissolved with apharmaceutically acceptable carrier, and then intravenouslyadministered.

In one aspect, provided herein is a method for the treatment ofbacterial infections in a mammal, comprising administering to saidmammal a therapeutically effective amount of a pharmaceuticalcomposition prepared according to the methods described herein. A methodfor the treatment of bacterial infections in a mammal can compriseadministering to said mammal a therapeutically effective amount of apharmaceutical composition comprising ceftolozane sulfate and sodiumchloride. Non-limiting examples of bacterial infections that can betreated by the methods of the invention include infections caused by:aerobic and facultative gram-positive microorganisms (e.g.,Staphylococcus aureus, Enterococcus faecalis, Staphylococcusepidermidis, Streptococcus agalactiae, Streptococcus pneumonia,Streptococcus pyogenes, Viridans group streptococci), aerobic andfacultative gram-negative microorganisms (e.g., Acinetobacter baumanii,Escherichia coli, Haemophilus influenza, Klebsiella pneumonia,Pseudomonas aeruginosa, Citrobacter koseri, Moraxella catarrhalis,Morganella morganii, Neisseria gonorrhoeae, Proteus mirabilis, Proteusvulgaris, Serratia marcescens, Providencia stuartii, Providenciarettgeri, Salmonella enterica), gram-positive anaerobes (Clostridiumperfringens), and gram-negative anaerobes (e.g., Bacteroides fragilisgroup (e.g., B. fragilis, B. ovatus, B. thetaiotaomicron, and B.vulgates), Bacteroides distasonis, Prevotella melaninogenica). Incertain embodiments of the methods described herein, bacterial infectionis associated with one or more of the following conditions: complicatedintra-abdominal infections, complicated urinary tract infections (cUTIs)and pneumonia (e.g., community-acquired, or nosocomial pneumonia).Community-acquired pneumonia (moderate severity only) can includeinfections caused by piperacillin-resistant, beta-lactamase producingstrains of Haemophilus influenza. Nosocomial pneumonia (moderate tosevere) caused by piperacillin-resistant, beta-lactamase producingstrains of Staphylococcus aureus and by Acinetobacter baumanii,Haemophilus influenzae, Klebsiella pneumoniae, and Pseudomonasaeruginosa.

As used herein, “treating”, “treat” or “treatment” describes themanagement and care of a patient for the purpose of combating a disease,condition, or disorder and includes the administration of apharmaceutical composition of the present invention to alleviate thesymptoms or complications of a disease, condition or disorder, or toreduce the extent of the disease, condition or disorder. The term“treat” can also include treatment of a cell in vitro or an animalmodel.

By a “therapeutically effective amount” of a compound of the inventionis meant a sufficient amount of the compound to treat the disorder(e.g., bacterial infection). The specific therapeutically effectiveamount that is required for the treatment of any particular patient ororganism (e.g., a mammal) will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound or composition employed; thespecific composition employed; the age, body weight, general health, sexand diet of the patient; the time of administration, route ofadministration, and rate of excretion of the specific compound employed;the duration of the treatment; drugs used in combination or coincidentalwith the specific compound employed; and like factors well known in themedical arts (see, for example, Goodman and Gilman's, “ThePharmacological Basis of Therapeutics”, Tenth Edition, A. Gilman, J.Hardman and L. Limbird, eds., McGraw-Hill Press, 155-173, 2001, which isincorporated herein by reference in its entirety). The therapeuticallyeffective amount for a given situation can be readily determined byroutine experimentation and is within the skill and judgment of theordinary clinician.

As used herein, “125-500 mg sodium chloride per 1000 mg of ceftolozane”refers to a ratio of sodium chloride to ceftolozane active. The phrase“125-500 mg sodium chloride per 1000 mg of ceftolozane” includes “62.5to 250 mg sodium chloride per 500 mg of ceftolozane active” and othersimilar weight ratios. In addition, “1,000 mg of ceftolozane asceftolozane sulfate” refers to an amount of ceftolozane sulfateeffective to provide 1,000 mg of ceftolozane. “Ceftolozane active refersto ceftolozane free base.” The amount of sodium per gram of ceftolozaneactivity in a pharmaceutical composition containing ceftolozane sulfateand sodium chloride can be calculated using the relevant molecularweights of ceftolozane, ceftolozane sulfate, sodium chloride and sodium.

As used herein, “1000 mg ceftolozane” refers to an amount of ceftolozanethat is considered a bioequivalent by the United States Food and DrugAdministration (FDA), i.e. for which 90% CI of the relative mean Cmax,AUC(0-t) and AUC(0-∝) is within 80.00% to 125.00% of the referenceformulation in the fasting state (see: “Guidance for Industry:Bioavailability and Bioequivalence Studies for Orally Administered DrugProducts—General Considerations”. Center for Drug Evaluation andResearch, United States Food and Drug Administration, 2003).

“Ceftolozane active” refers to the active portion of a salt form ofceftolozane, the free base form of ceftolozane.

As used herein, the term “tazobactam active” refers to the activeportion of a salt form of tazobactam, tazobactam free acid.

In another aspect, the disclosed sodium chloride stabilized ceftolozanecompositions can be characterized by decrease in ceftolozane totalpurity is not greater than 3.7% after storing the pharmaceuticalcomposition for seven days (e.g., in a sealed container) at 60° C., asdetermined by HPLC using a Develosil column ODS-UG-5; 5 micrometers;250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of45° C. In another aspect, the disclosed sodium chloride stabilizedceftolozane compositions can be characterized by decrease in ceftolozanetotal purity is not greater than 4.2% after storing the pharmaceuticalcomposition for seven days at 60° C., as determined by HPLC using aDevelosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase ofsodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0mL/min flow rate and oven temperature of 45° C. In another aspect, thedisclosed sodium chloride stabilized ceftolozane compositions can becharacterized by decrease in ceftolozane total purity is not greaterthan 4.5% after storing the pharmaceutical composition for seven days(e.g., in a sealed container) at 60° C., as determined by HPLC using aDevelosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase ofsodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0mL/min flow rate and oven temperature of 45° C. In another aspect, thedisclosed sodium chloride stabilized ceftolozane compositions can becharacterized by decrease in ceftolozane total purity is not greaterthan 5.0% after storing the pharmaceutical composition for seven days(e.g., in a sealed container) at 60° C., as determined by HPLC using aDevelosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase ofsodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0mL/min flow rate and oven temperature of 45° C. In another aspect, thedisclosed sodium chloride stabilized ceftolozane compositions werecharacterized by an increase in the amount of the impurity representedby Peak 1 not greater than 1.8% after storing the pharmaceuticalcomposition for seven days (e.g., in a sealed container) at 60° C., asdetermined by HPLC using a Develosil column ODS-UG-5; 5 micrometers;250×4.6 mm, a mobile phase of sodium perchlorate buffer solution (pH2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oven temperature of45° C., where Peak 1 has a retention time relative to ceftolozane of0.1. In another aspect, the disclosed sodium chloride stabilizedceftolozane compositions were characterized by an increase in the amountof the impurity represented by Peak 1 not greater than 2.0% afterstoring the pharmaceutical composition for seven days (e.g., in a sealedcontainer) at 60° C., as determined by HPLC using a Develosil columnODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodiumperchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/minflow rate and oven temperature of 45° C., where Peak 1 has a retentiontime relative to ceftolozane of 0.1. In another aspect, the disclosedsodium chloride stabilized ceftolozane compositions were characterizedby an increase in the amount of the impurity represented by Peak 1 notgreater than 2.2% after storing the pharmaceutical composition for sevendays (e.g., in a sealed container) at 60° C., as determined by HPLCusing a Develosil column ODS-UG-5; 5 micrometers; 250×4.6 mm, a mobilephase of sodium perchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v)at a 1.0 mL/min flow rate and oven temperature of 45° C., where Peak 1has a retention time relative to ceftolozane of 0.1.

ILLUSTRATIVE EXAMPLES OF SELECTED EMBODIMENTS OF THE INVENTION Example 1Manufacturing Procedure of Bulk (Tray) Lyophilized Ceftolozane

There are four main steps in the manufacture of CXA-101 bulk drugproduct: dissolution, sterile filtration, bulk lyophilization, andpackaging into Sterbags®. These four main steps are composed of a totalof 20 minor steps. The CXA-101 bulk drug product manufacturing processis presented below.

I. Dissolution

1. The prescribed amount of water for injection (“WFI”) is charged intothe dissolution reactor.

2. A prescribed amount of citric acid is added.

3. The solution is cooled at 5° C. to 10° C.

4. A prescribed amount of CXA-101 drug substance is added to thesolution.

5. A prescribed amount of L-arginine is slowly added to the solution.

6. A check for complete dissolution is performed. Solution pH isverified to be in the target range of 6.5 to 7.0.

7. A prescribed amount of sodium chloride is added to the solution.

8. A check for complete dissolution is performed. Solution pH isverified to be in the target range of 6.0 to 7.0. If the pH is out ofthis range adjust with either L-Arginine or citric acid.

9. WFI is added to bring the net weight to 124.4 kg and the solution ismixed well.

10. Samples are withdrawn for testing of final pH.

II. Sterile Filtration

11. The solution is passed through the filter (pore size 0.45 μm)followed by double filters (pore size 0.22 μm) onto a shelf on theCriofarma lyophilizer.

12. The line is washed with WFI.

13. The washing solution is passed from Step 12 through sterilefiltration.

III. Bulk Lyophilization

14. The washing solution is loaded onto a separate shelf in thelyophilizer (and later discarded).

15. The solution is lyophilized until dry.

16. The product shelf is cooled to 20° C.±5° C.

IV. Packaging into Sterbags®

17. The lyophilized bulk drug product powder is milled.

18. The milled powder is sieved.

19. The sieved powder is blended for 30 minutes.

20. The powder is then discharged into Sterbags®

Prefiltration and Sterile-Filtration

Filtrate the compounded solution with a sterile tilter-set whichconsists of a 0.2 um polyvinylidene fluoride membrane filter (Durapore®,Millipore) and a 0.1 um polyvinylidene fluoride membrane filter(Durapore®, Millipore) connected in tandem. Confirm the integrity ofeach filter before and after the filtration. Take approximately 100 mLof the filtrate in order to check bioburden.

Filter the prefiltered compounded solution through a sterile filter-setwhich consists of a 0.2 um polyvinylidene fluoride membrane filter and a0.1 um polyvinylidene fluoride membrane filter connected in tandem, andintroduce the final filtrate into an aseptic room. Confirm the integrityof each filter before and after the filtration.

Processing of Vial, Stopper and Flip-Off Cap

Wash a sufficient quantity of 28 mL vials with water for injection andsterilize the washed vials by a dry-heat sterilizer. Then transfer thesterilized vials into a Grade A area located in an aseptic room.

Wash a sufficient quantity of stoppers with, water for injection.Sterilize and dry the washed stoppers by steam sterilizer. Then transferthe sterilized stoppers into a Grade A area located in an aseptic room.

Sterilize a sufficient quantity of flip-off caps by steam sterilizer.Then transfer the sterilized flip-off caps into a Grade A or B arealocated in an aseptic room.

Filling and Partially Stoppering

Adjust the fill weight of the filtered compounded solution to 11.37 g(corresponds to 10 mL of the compounded solution), then start fillingoperation. Check the filled weight in sufficient frequency and confirmit is in target range (11.37 g±1%, 11.26 to 11.43 g). When deviationfrom the control range (11.37 g±2%, 11.14 to 11.59 g) is occurred,re-adjust the filling weight.

Immediately after a vial is filled, partially stopper the vial with asterilized stopper. Load the filled and partially stoppered vials ontothe shelves of a lyophilizer aseptically.

Lyophilization to Crimping, Visual Inspection, Labeling and Packaging

After all filled and partially stoppered vials are loaded into alyophilizer, start the lyophilization program shown in FIG. 2. Freezethe loaded vials at −40° C. and keep until all vials freeze. Forward theprogram to primary drying step (shelf temperature; −20° C., chamberpressure; 100 to 150 mTorr). Primary drying time should be determined bymonitoring the product temperature. Forward the program to secondarydrying step (shelf temperature; 30° C., chamber pressure; not more than10 mTorr) after completion of the primary drying step. After all vialsare dried completely, return the chamber pressure to atmosphericpressure with sterilized nitrogen. Then stopper vials completely.

Unload the lyophilized vials from the chamber and crimp with sterilizedflip-off caps.

Subject all crimped vials to visual inspection and label and package allpassed vials.

Example 2 Analytical HPLC Method A. Operative Conditions

-   Column Develosil ODS-UG-5; 5 μm, 250×4.6 mm (Nomura Chemical, Japan)-   Mobile phase Sodium Perchlorate Buffer Solution (PH 2.5)/CH₃CN 90:10    (vlv)-   Flow rate 1.0 mL/min-   Wavelength 254 nm-   Injection volume 10 μL-   Oven Temperature 45° C.-   Run Time 85 minutes

Gradient Profile:

Time (min) A % B % 0 75 25 30 70 30 60 0 100 85 0 100 85.1 75 25 110 7525

B. Mobile Phase Preparation.

Sodium Perchlorate Buffer Solution was made by dissolving 14.05 g ofsodium perchlorate Monohydrate in 1000.0 mL of water followed byadjusting pH to 2.5 with diluted perchloric acid (1 in 20).

Mobile Phase was then made by mixing Sodium Perchlorate Buffer Solution(pH 2.5) and acetonitrile in the ratio 90:10 (v/v).

Sodium Acetate Buffer Solution pH 5.5 (Diluent) was made by dissolving1.36 g of sodium acetate trihydrate in 1000.0 mL of water followed byadjusting to pH 5.5 with diluted acetic acid (1 in 10).

C. Sample Preparation.

Sample solution: dissolve 20.0 mg, exactly weighed, of Sample, in 20.0mL of water (Prepare just before injection into HPLC system).

System Suitability Solution (1%): take 1.0 mL of the Sample Solution(use first sample if more are present) and transfer into a 100.0 mLvolumetric flask, dilute with water to volume and mix.

D. HPLC Analysis Procedure

-   1. Inject Blank (water)-   2. Inject System Suitability Solution and check for tailing factor    and theoretical plate number for CXA-101 peak:    -   The tailing factor must not be greater than 1.5    -   Theoretical plates number must not be less than 10000-   3. Inject Sample Solution-   4. Inject System Suitability Solution and check for tailing factor    and theoretical plate number for CXA-101 peak.    -   The tailing factor must not be greater than 1.5    -   Theoretical plates number must not be less than 10000-   5. Identify the peaks of Related Substances in the Sample    chromatogram based on the reference chromatogram reported in FIGS.    1A and 1B or, alternatively, on the basis of the RRT values reported    in Table 1 (FIG. 3). The material for “Peak 1” in the Table 1 of    FIG. 3, with a retention time of about 0.14 relative to ceftolozane    is believed to have the chemical structure of formula (II):

E. Calculations

-   I. Report for each related substance its amount as expressed by area    percent.

$C_{i} = \frac{A_{i} \times 100}{A_{t} + {\Sigma \; A_{i}}}$

wherein:

C_(i)=Amount of related substance i in the Sample, area %

A_(i)=Peak area of related substance i in the Sample chromatogram

A_(t)=Area of CXA-101 peak in the Sample chromatogram

A_(t)+Σ A_(i)=Total peaks area in the Sample chromatogram

Consider as any Unspecified Impurity, each peak in the chromatogramexcept CXA-101, peaks from 1 to 11 and every peak present in the blankchromatogram and report the largest.

-   II. Report the total impurities content as expressed by the    following formula:

$C_{T} = \frac{A_{i} \times 100}{A_{t} + {\Sigma \; A_{i}}}$

wherein:

C_(T)=total impurities content in the Sample, area %

A_(t)=area of CXA-101 peak in the sample chromatogram

Σ A_(i)=total peak areas of impurities in the sample chromatogram

Example 3 The Amount of Sodium Chloride can be Selected to StabilizeCeftolozane in Pharmaceutical Compositions

A 7-day stability study of multiple ceftolozane pharmaceuticalcompositions was carried out at 60% RH in accordance with ICHguidelines. This stability study examined the effect of temperature andhumidity on various ceftolozane pharmaceutical compositions bulk drugproduct stability when stored in the container closure configuration ofSterbag®.

The amount of ceftolozane in ceftolozane sulfate can similarly becalculated based on the respective molecular molar weights ofceftolozane and ceftolozane sulfate (e.g., 1,147 mg ceftolozane sulfatecontains about 1,000 mg of ceftolozane). Accordingly, a compositioncomprising about 1,147 mg ceftolozane sulfate and 125-500 mg of sodiumchloride also contains 125-500 mg of sodium chloride per 1,000 mg ofceftolozane.

A. CXA-101 Purity Increases in Compositions having at Least About 125 mgNaCl/1,000 mg Ceftolozane Active

A stability study was carried out at 60° C. as described in Example 3.Sodium chloride content in test samples is described in Table 2 (FIG.4). The samples were formulated with 481, 190, 125, 75, and 50 mg sodiumchloride per lg of ceftolozane active.

FIG. 5A is table 3 with data for total purity of ceftolozane measured byHPLC during the 7-day stability test using the HPLC method in Example 2,with the data plotted in the graph of FIG. 5B.

B. The Amount of Substances Identified by HPLC Peaks 1 and 7 Decreasesin Compositions having at Least About 125 mg NaCl/1,000 mg CeftolozaneActive

A stability study was carried out at and 60° C. as described in Example3. Sodium chloride content in test samples is described in Table 2 (FIG.4). The samples were formulated with 481, 190, 125, 75, and 50 mg sodiumchloride per lg of ceftolozane active.

Stability data for amounts of additional substances in the ceftolozanecompositions from Table 2 (FIG. 4) as measured by peaks 1, 3 and 7 byHPLC (according to Example 2) are summarized in Tables 4-6, FIGS. 6A, 7Aand 8A. The data are also plotted in FIGS. 6B, 7B, and 8B to show trendsof total purity, peak 1, RRT 0.43+peak 3, and peak 7 with respect tosodium chloride (NaCl), respectively. FIG. 5A is Table 3 with totalpurity of ceftolozane measured by HPLC during the 7-day stability test,with the data plotted in the graph of FIG. 5B.

Example 4 The Amount of Sodium Chloride can be Selected to StabilizeCeftolozane in Pharmaceutical Compositions Comprising Ceftolozane andTazobactam

A. Stabilized Ceftolozane Compositions Co-Lyophilized with Tazobactam

A composition comprising ceftolozane, sodium chloride and tazobactam wasprepared by co-lyophilizing a stabilizing amount of sodium chloride,ceftolozane sulfate and tazobactam acid in amounts described in Table 7(FIG. 9A) together in an aqueous solution to obtain a stabilizedceftolozane lyophilized composition (“Co-Lyophilized Combo DrugProduct”). The components of the composition that was lyophilized toobtain the Co-Lyophilized Combo Drug Product is shown in Table 7 (FIG.9A). Notably, this composition included about 484 mg of sodium chlorideper 1,000 mg ceftolozane active provided as ceftolozane sulfate, and aweight ratio of about 2:1 between the CXA101 and tazobactam acid.

The results of a stability study of the Co-Lyophilized Combo DrugProduct are shown in Table 8 (FIG. 9B) and Table 9 (FIG. 9C) asrepresentative examples that summarize the results at 25° C./RH=60% and40° C./RH=75% after one month (T1) and three months (T2). Samples wereanalyzed using a HPLC method as described in Example 2.

Referring to the data in Tables 8 and 9, the Co-Lyophilized Combo DrugProduct was characterized by amounts of the substances corresponding toHPLC peaks 1-12 that were less than the applicable drug productspecification, indicating stabilization of the ceftolozane in thepresence of about 484 mg of sodium chloride per 1,000 mg of ceftolozaneactive.

B. Stabilized Ceftolozane Compositions Lyophilized without Tazobactam

A composition comprising ceftolozane, and a stabilizing amount of sodiumchloride was lyophilized, and then blended with a separately lyophilizedcomposition of tazobactam. The stabilized ceftolozane composition wasformed by lyophilizing an aqueous solution of the “CXA-101 for InjectionBulk” row of Table 10 (FIG. 10A), which was then blended with 5.4 gtazobactam free acid to form a pharmaceutical composition containingceftolozane, sodium chloride and tazobactam components (“Blend ComboDrug Product”). Notably, this composition included about 481 mg ofsodium chloride per 1,000 mg ceftolozane active provided as ceftolozanesulfate, and a weight ratio of about 2:1 between the CXA101 andtazobactam acid.

The results of a stability study of the Blend Combo Drug Product areshown in Table 11 (FIG. 10B) and Table 12 (FIG. 10C) as representativeexamples that summarize the results at 25° C./RH=60% and 40° C./RH=75%after one month (T1) and three months (T2). Samples were analyzed usinga HPLC method as described in Example 2.

Referring to the data in tables 11 and 12, the Blend Combo Drug Productwas characterized by amounts of the substances corresponding to HPLCpeaks 1-12 that were less than the applicable drug productspecification, indicating stabilization of the ceftolozane in thepresence of about 481 mg of sodium chloride per 1,000 mg of ceftolozaneactive.

Example 6 Improvement in the Purity of Ceftolozane CXA-201PharmaceuticalCompositions with Varying Amounts of Sodium Chloride

A stability study was carried out at 60° C. as described in Example 3.The sodium chloride content in the CXA-201 compositions is described inTable 13, FIG. 11. The HPLC data at 60° C. are summarized in Table14-17, FIGS. 12-15.

What is claimed is:
 1. A pharmaceutical composition comprising 125 mg to500 mg sodium chloride per 1,000 mg of ceftolozane active, wherein thedecrease in ceftolozane total purity is not greater than about 4% afterstoring the pharmaceutical composition for seven days in a sealedcontainer at 60° C., as determined by HPLC using a Develosil columnODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodiumperchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/minflow rate and oven temperature of 45° C.
 2. The pharmaceuticalcomposition of claim 1, further comprising L-arginine.
 3. Thepharmaceutical composition of claim 1, further comprising citric acid.4. The pharmaceutical composition of claim 2, further comprising citricacid.
 5. The pharmaceutical composition of claim 1, wherein thepharmaceutical composition is formulated for parenteral administration.6. The pharmaceutical composition of claim 5, wherein the composition isa unit dosage form in a vial comprising 125 mg to 500 mg sodiumchloride, 1,000 mg of ceftolozane in the form of ceftolozane sulfate,L-arginine and citric acid.
 7. The pharmaceutical composition of claim1, wherein the pharmaceutical composition is lyophilized.
 8. Thepharmaceutical composition of claim 1, wherein the ceftolozane isceftolozane sulfate.
 9. A pharmaceutical composition comprising 125 mgto 500 mg sodium chloride per 1,000 mg of ceftolozane active, whereinthe increase in the amount of the impurity represented by Peak 1 is notgreater than about 2% after storing the pharmaceutical composition forseven days at 60° C., as determined by HPLC using a Develosil columnODS-UG-5; 5 micrometers; 250×4.6 mm, a mobile phase of sodiumperchlorate buffer solution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/minflow rate and oven temperature of 45° C., where Peak 1 has a retentiontime relative to ceftolozane of about 0.1.
 10. The pharmaceuticalcomposition of claim 9, further comprising L-arginine.
 11. Thepharmaceutical composition of claim 9, further comprising citric acid.12. The pharmaceutical composition of claim 10, further comprisingcitric acid.
 13. The pharmaceutical composition of claim 9, wherein thepharmaceutical composition is formulated for parenteral administration.14. The pharmaceutical composition of claim 13, wherein the compositionis a unit dosage form comprising 125 mg to 500 mg sodium chloride, 1,000mg of ceftolozane in the form of ceftolozane sulfate, L-arginine andcitric acid.
 15. The pharmaceutical composition of claim 9, wherein thepharmaceutical composition is lyophilized.
 16. The pharmaceuticalcomposition of claim 9, wherein the ceftolozane is ceftolozane sulfate.17. A unit dosage form injectable pharmaceutical composition comprising125 mg to 500 mg sodium chloride and 1,000 mg of ceftolozane activepresent as a composition of formula (I)


18. A solid pharmaceutical composition comprising 125 mg to 500 mgsodium chloride per 1,000 mg of ceftolozane active present asceftolozane sulfate, wherein the ceftolozane total purity is at leastabout 94% after storing the pharmaceutical composition for three days at60° C., as determined by HPLC using a Develosil column ODS-UG-5; 5micrometers; 250×4.6 mm, a mobile phase of sodium perchlorate buffersolution (pH 2.5)/CH₃CN 90:10 (v/v) at a 1.0 mL/min flow rate and oventemperature of 45° C.